In Vitro and In Vivo Micronucleus Tests in Genotoxicological Research

U današnje vrijeme životni i radni okoliš čovjeka nepredvidiva je mješavina kemijskih spojeva od kojih neki mogu djelovati kao mutageni ili kao aneugeni karcinogeni. Osim toga, razni su izvori zračenja dodatno opterećenje na genom čovjeka. Od desetak metoda koje se rabe u genotoksikološkim istraživanjima, in vitro i in vivo mikronukleus tehnike posljednjih su godina našle primjenu kako u bazičnim tako i u kliničkim i farmakološkim istraživanjima. Prednost in vitro i in vivo mikronukleus tehnika je da istodobno pružaju podatke o klastogenom i aneugenom djelovanju agenasa. Kako ne zahtijeva kultiviranje stanica, in vivo mikronukleus tehnika, iako uvedena nekoliko godina nakon in vitro tehnike, zbog svoje jednostavnosti i brzine primjene postaje sve važniji izvor podataka u akutnim i kroničnim genotoksikološkim istraživanjima.Working and living environment today is an unpredictable mixture of different chemical substances. Some of these chemicals are mutagenic carcinogens and others are aneugenic carcinogens. An additional burden to the human genome is the exposure to several kinds of radiations. Among a dozen genotoxicological methods in use in vitro and in vivo, the micronucleus assay has found application in the basic research, clinical reasearch, and pharmacological studies. The advantage of in vivo and in vitro micronucleus assays is in the fact that they provide data on both clastogenic and aneugenic action of agents. Although several years behind the in vitro technique, the in vivo micronucleus assay has swiftly developed into a significant source of data in acute and chronic genotoxic investigations, as it does not require cell culture

Anemia and inflammatory bowel diseases

Cilj istraživanja: Cilj ovog istraživanja bio je utvrditi učestalost i vrstu anemije u odraslih osoba oboljelih od upalnih bolesti crijeva s područja istočne Hrvatske te utvrditi postoji li povezanost između pojavnosti anemije te vrijednosti C-reaktivnog proteina u krvi oboljelih kao neizravnog pokazatelja aktivnosti upale. Ispitanici i metode: Provedeno istraživanje ustrojeno je po načelu cross-sectional studije odnosno kao tipična metoda istraživanja presjeka ili prevalencije. U istraživanje je uključeno ukupno 78 oboljelih od upalnih bolesti crijeva s područja istočne Hrvatske u rasponu od 22 do 69 godina. Korištene su sljedeće laboratorijske analize za dokazivanje anemije: određivanje željeza u serumu, transferina, feritina, TIBC-a,UIBC-a, C-reaktivnog proteina, kompletne krvne slike i retikulocita. Rezultati: Ovim istraživanjem utvrđena je prisutnost anemije kod 24,4 % ispitanika. Prema vrsti anemije, u 63,2 % slučajeva riječ je o sideropeničnoj anemiji, a u 36,8 % slučajeva riječ je o anemiji kronične bolesti. Nema statistički značajne razlike u učestalosti i vrsti anemije između oboljelih od Crohnove bolesti te oboljelih od ulceroznog kolitisa. Kod osoba s aktivnom bolešću statistički je značajno češće utvrđeno postojanje anemije u odnosu na osobe s inaktivnom bolešću. Zaključak: Anemija je značajna ekstraintestinalna komplikacija upalnih bolesti crijeva te kako slijedom izostanka liječenja anemije u sklopu upalne bolesti crijeva anemija predstavlja značajan dodatni simptom odnosno komplikaciju kod oboljelih.Objectives: The aim of this study was to determine the frequency and type of anaemia in adult patients with inflammatory bowel disease in the Eastern Croatia. Furthermore, the aim was to examine whether there is correlation between the incidence of anaemia and the value of C-reactive protein in patients’ blood, which could be considered an indirect indicator of the inflammation. Participants and methods: The study is organized as a cross-sectional study, i. e. cross-section and prevalence survey method. The study included 78 patients with IBD from Eastern Croatia 22 – 69 years of age. The following laboratory analyses for the detection of anaemia were used: serum levels of iron, transferrin, ferritin, TIBC, UIBC, C-reactive protein, complete blood count and reticulocyte count. Results: comorbidity present in 24.4% of patients with inflammatory bowel disease in Eastern Croatia. Anaemia was present in 24.4% of the subjects. According to the type of anaemia, in 63.2% of cases it is sideropenic anaemia, and in 36.8% of cases the chronic disease anaemia. Statistically significant differences were not found in the frequency and type of anaemia in patients with Crohn's disease in relation to patients with ulcerative colitis. In patients with active illness, there is a statistically significant increase in the presence of anaemia in relation to patients with inactive disease. Conclusion: Anaemia is a significant extra-intestinal complication of inflammatory bowel disease and in the absence of the treatment of anaemia within the inflammatory bowel disease it is a significant additional symptom or complication in the affected population

Traumatske hemolitične anemije: dva prikaza slučaja i pregled literature

Two patients with intravascular hemolysis are presented. One patient had traumatic hemolytic anemia after mechanical heart valve reimplantation, and the diagnosis was made rapidly and easily. In the other patient, the diagnosis was not so easy complex and time-consuming. This patient had both, traumatic hemolytic anemia caused by orthopedic prosthesis, and neutropenia. Fragmentation was caused by interaction of red blood cells with altered intravascular surfaces or by direct physical trauma to the cells due to excessive shear forces in the circulation. The following findings are seen in intravascular hemolysis: decreased hemoglobin, increased reticulocytes and lactic dehydrogenase, decreased serum haptoglobin, and presence of shizocytes in peripheral blood smear. Patients are considered to have intravascular hemolysis when lactic dehydrogenase is >460, along with the presence of two established criteria. If not causal, therapy is supportive.Prikazana su dvojica bolesnika s intravaskularnom hemolizom, od kojih je jedan imao hemolitičnu anemiju nakon ponovljene ugradnje mehaničkog srčanog zalistka i u kojega je dijagnoza bila brza i jednostavna. U drugoga je bolesnika postavljanje dijagnoze zahtijevalo dosta vremena i truda. Ovaj je bolesnik imao mehaničku hemolitičnu anemiju uzrokovanu ortopedskim protezama, ali i neutropeniju. Fragmentacija je bila izazvana interakcijom eritrocita s promijenjenom površinom žile ili izravnom fizikalnom traumom stanica u krvotoku. Kod intravaskularne hemolize prisutni su sljedeći nalazi: sniženi hemoglobin, povišeni retikulociti i laktat dehidrogenaza, sniženi haptoglobin u serumu, te prisutnost shizocita u razmazu periferne krvi. Smatra se da bolesnici imaju intravaskularnu hemolizu kada je laktat dehidrogenaza viša od 460, uz prisutnost dvaju utvrđenih kriterija. Ako nije etiološka, terapija je potporna

The assessment of DNA damage and repair following exposure to sevoflurane in vivo

Cilj istraživanja bio je istražiti povezanost oštećenja i popravka DNA u leukocitima periferne krvi, stanicama jetre, bubrega i mozga Swiss albino miševa uzrokovanog uzastopnim izlaganjem sevofluranu. Miševi su bili izlagani sevofluranu u dozi od 2.4 vol% po dva sata dnevno, 3 dana uzastopce. Za procjenu genetičkog oštećenja spomenutih stanica upotrijebljeni su komet test i mikronukleus test. Za provođenje komet testa miševi su podijeljeni u 5 pokusnih skupina (kontrolna skupina, skupina miševa žrtvovanih treći dan pokusa neposredno nakon izlaganja sevofluranu, 2 sata, 6 sati i 24 sata nakon izlaganja sevofluranu). Rezultati su pokazali statistički značajno povećanje srednje vrijednosti TL, TM i TI u svim pregledanim organima u usporedbi sa kontrolom. Veličina oštećenja DNA u točno određenom vremenu nakon uzastopnog izlaganja sevofluranu, bila je drugačija za svaki pregledani organ. Najveće oštećenje DNA uočeno je odmah nakon izlaganja sevofluranu, posebno u leukocitima periferne krvi. Inducirano oštećenje se dogodilo šest sati kasnije u organima nego u stanicama krvi, što je bilo za očekivati zbog toksikokinetike lijekova, jer se sevofluran prvo apsorbira u krvi. Niti jedno od proučavanih tkiva nije pokazivalo znakove popravka sve dok nisu prošla 24 sata od izlaganja sevofluranu. Za detektiranje nepopravljenih oštećenih dijelova genoma in vivo upotrijebljen je mikronukleus test. Broj mikronukleusa u retikulocitima statistički se značajno povećao u svim proučavanim vremenskim razmacima nakon tretmana u odnosu na kontrolnu skupinu.Aim of the study was to investigate the relationship between DNA damage and repair of peripheral blood leucocytes (PBL), liver, kidney and brain cells of Swiss albino mice induced by repeated exposure to sevoflurane. Mice were exposed to sevoflurane in a dose of 2.4vol% for two hours daily, for three consecutive days. Genetic damage of mentioned cells was investigated using comet assay and micronucleus test. To perform comet assay, mice were divided into 5 experimental groups (control group, groups of exposed mice sacrificed third day of experiment immediately after exposure ot sevoflurane, two hours, six hours and twenty-four hours later). The results showed statistically significant increase in mean TL, TM and TI values in all examined organs vs. the control group. The DNA damage at a specific time after the repeated exposure to sevoflurane was different for each examined organ. Significant DNA damage immediately post exposure to sevoflurane was observed in PBL. The induced damage in organs occured 6 hours later than in PBL what was expected according to toxicokinetic of drugs, where blood is the first compartment to absorb the sevoflurane. However none of the assayed tissues revealed signs of repair until 24th hour after the exposure. To distinguish unrepaired genome damage in vivo micronucleus test was applied. Number of micronuclei in reticulocytes showed statistically significant increase compared with control group at all observed times after the treatment

The analysis of the chromosomal aberrations frequency of the employees from different workplaces in the zone of ionizing radiation.

Cilj predstavljenog istraživanja je da ispita uticaj malih doza jonizujućeg zračenja na zaposlene na različitim radnim mestima u zoni jonizujućeg zračenja. Materijal i metode: Za studiju retrospektivne prirode analizirani su rezultati dobijeni iz kartona preventivno-periodičnih medicinskih pregleda 148 zaposlenih na različitim radnim mestima u zoni jonizujućeg zračenja u Kliničkom Centru Srbije a koji obavljaju preglede u Institutu za medicinu rada Srbije. Ispitanici su podeljeni u tri velike grupe u odnosu na izvor zračenja-oblast rada na zaposlene u nuklearnoj medicini, intrerventnoj radiologiji i opštoj radiologiji. Podaci korišćeni za studiju su podaci sa prethodnog/prvog periodičnog pregleda koji je obavljan pre ulaska u zonu jonizujućeg zračenja ili u toku prve godine izlaganja (u toku obavljanja specijalizacije, u skladu sa odlukom poslodavca da pošalje zaposlenog na pregled) i podaci sa poslednjeg periodičnog pregleda. Uzimani su u obzir ukupni i ekspozicioni radni staž, vrsta izvora zračenja, hematološki parametri (eritrociti, retikulociti, leukociti sa leukocitarnom formulom, trombociti), frekvenca i broj nestabilnih hromozomskih aberacija (ishod testa nestabilnih hromozomskih aberacija) kao i ishod mikronukleusnog testa i njihov broj (značajnih za ocenu radne sposobnosti), primljena kumulativna petogodišnja doza zračenja, morbiditet ispitanika i prisutvo profesionalnih bolesti. Hematološki parametri (eritrociti-Er, retikulociti-Rtc, ukupni leukociti-Leu, monociti-Mo, neutrofili-Neut, eozinofili-Eo, bazofili-Ba, limfociti-Ly i trombociti-Tr) su dobijani ispitivanjem periferne venske krvi u aparatu Beckman Coulter HMX ( na poslednjem periodičnom pregledu) a na prethodnom su uglavnom ispitivani starom metodom ručnog brojanja ćelija (preračunatih za statistički obradu). Nestabilne hromozomske aberacije su dobijane modifikovanom Moorhead’s mikro metodom. Mikronukleusni test je rađen metodom po Fenech-u i Morley-u...The aim of the research is to analyse the influence of the small doses of ionizing radiation on the employees from different workplaces in the zone of ionizing radiation. Material and methods: For this study of the retrospective nature we have analyzed results obtained from the medical records based on preventive-periodical examinations of 148 persons employed at different workplaces in the zone of ionizing radiation at the Clinical Center of the Republic of Serbia. All of the employees go to medical examinations at the Institute of Occupational Medicine of Serbia. The employees were divided into three large groups as related to the source of radiation – the field of work: the nuclear medicine employees, the interventional radiology employees and the general radiology employees. The data was taken from the previous/initial checkup which had been done before the entrance to the zone of ionizing radiation or during the first year of exposure to radiation (during the specialization period, in accordance with employer’s decision to send an employee to the examination) as well as the data from the last periodical examination. We also took into consideration total and exposition work periods, type of radiation source, haematological parameters (erythrocytes, reticulocytes, leukocytes with differential blood count, platelets), frequency and number of unstable chromosomal aberrations (the results of the test of the unstable chromosomal aberrations), the results of micronucleus test and their number (important for the assessment of working ability), as well as received cumulative five-year radiation dose, morbidity of patients and presence of occupational diseases. The hematological parameters (erythrocytes-RBC, reticulocytes-Rtc, total leucocytes- WBS, monocytes-Mo, neutrophils-Neu, eosinophils-Eo, basophils-Ba and platelets) were obtained through processing of the peripheral vein blood by Beckman Coulter HMX apparatus (at the periodical checkup), while at the previous / initial checkup it was done through the old method of manual cell counting (recalculated for statistical processing)..

Long-Term Bone Marrow Culture

Dugotrajna kultura koštane srži eksperimentalni je model hematopoeze koji in vitro reproducira uvjete hematopoetskog mikrookružja in vivo. U dugotrajnoj kulturi postiže se proliferacija i diferencijacija primitivnih matičnih hematopoetskih stanica mezenhimskog podrijetla. Dugotrajnu kulturu koštane srži čine adherentni stromalni sloj i slobodne stanice u tekućem dijelu (supernatantu) kulture. U dugotrajnoj se kulturi hematopoeza zbiva bez egzogenih poticatelja rasta jer njih stvara stromalni sloj. Izgled i razvitak strome, proliferacija i diferencijacija progenitorskih stanica i proizvodnja citokina razlikuju se u dugotrajnoj kulturi normalne i patološki promijenjene koštane srži. Dugotrajna se kultura koštane srži upotrebljava u fundamentalnim istraživanjima hematopoeze, u farmakološkim ispitivanjima učinaka lijekova i imunocitokina na hematopoezu, za čišćenje autotransplantata koštane srži od preostalih leukemijskih stanica te za prijenos gena. Također je prikladan laboratorijski model za in vitro ispitivanje karcinogenih i drugih štetnih činitelja iz okoline koji utječu na krvotvorno tkivo pri profesionalnom ili habitualnom izlaganju.Long-term bone marrow culture is an experimental in vitro model of hematopoiesis imitating conditions in vivo. It contains hematopoietic elements at various stages of differentiation as well as a supportive stromal microenvironment. Primitive hematopoietic stem cells of mesenchymal origin, the long-term culture initiating cells proliferate and differentiate into different cell types, giving rise to the adherent stromal layer and to various hematopoietic elements attached to it or floating freely in the supernatant medium. The stromal layer keeps the hematopoietic cells aggregated, helps their mitosis, differentiation and maturation by cell-to-cell contact, produces hematopoietic growth factors (cytokines), and forms the extracellular matrix required for cell attachment. Hematopoiesis occurs without exogenous growth factors. The appearance and development of the stroma, the proliferation and differentiation of hematopoietic progenitor cells, and the production of cytokines differ in long-term cultures of the normal and of pathologically altered bone marrow. Long-term bone marrow culture is applied in fundamental studies of normal and pathologically altered hematopoiesis, in pharmacological research, in the purging of residual leukemia cells from bone marrow autotransplants, and in the gene transfer. It is also suitable for testing carcinogenic and toxic chemicals causing hematopoietic damage through occupational or habitual exposure

FAST AS ONE SPIRITUAL DIET

Uvod: Post predstavlja suzdržavanje od hrane i pića određeni dio vremena. Tu može biti riječ o “ukidanju” određenih vrsta hrane na neko određeno vrijeme ili apsolutno suzdržavanje od hrane i pića jedan cijeli dan ili čak nekoliko dana. Post ima veliku religioznu važnost (za različite vjernike), ali je važan i za zdravlje, ako ga se provodi pravilno. Duhovna kondicija /duhovna spremnost-duhovna spremnina nije duhovno stanje per se već ono uveliko ovisi o primjeni razumijevanja naše tjelesne spremnosti na duhovnu dimenziju. Cilj: Cilj ovog osvrta predmnijeva da bi se post mogao shvatiti i stanovitom vrstom duhovne dijete pače i određenom vrstom «duhovne tablete». Metode: Analizirani su anamnestičkih podataka o navikama i fiziološkim funkcijama poput onih o vjeri/svjetonazoru/stilu života odn. o apetitu /ritmu uzimanja obroka, njihovoj količini i kakvoći dobivenih u intervjuima prije uključivanja u bilo kakav psihijatrijski/psihoterapijski ili hagioterapijski tretman. Rezultati: Nešto točnije podatke o postu je jedino bilo moguće izvući iz podataka onih koji su se izjašnjavali vjernicima a koja su se odnosila na njihova prisustvovanja i neprisustvovanja propisanim religijskim obredima tj. prakticiranjima vjere odn. njezinih praksi iz pobožnosti. Omjer « redovitog prakticiranja» i «neredovitog prakticiranja» bi se mogao držati nesigurnim obzirom na poprilični raskorak podataka o manje ili više aktivne nesklonosti propisanoj religijskoj praksi u odnosu na samu vjersku privrženost. Izgleda da bi jedna od dilema liječnika, napose onih iz oblasti javnog zdravlja / poglavito onih koji se k tome bave i hagioterapijom mogla biti slijedeća: Kako dati opće prihvatljive preporuke, što je to zadovoljavajući ( povremeni, kratkoročni post ) te kakav bi trebao biti optimalan dugoročni, produženi post koji ne bi štetio u konkretnom kliničkom ili izvan bolničkom tretmanu posebice određenih populacijskih skupina poput mladih u razvoju ili starijih osoba. Zaključak: Ipak, unatoč navedenog raspravnog sadržaja, post kako onaj povremeni, kratkoročni tako ( donekle ) i onaj tzv. optimalni dugoročni valja promatrati kao i mogući resurs za unapređenje zdravlja i prevenciju bolesti ali i kao možebitni učvršćivač postojećeg, a onda i kao jednu od mjera koja bi mogla zamjetno pozitivno utjecati na duhovno zdravlje populacije ( učvršćivanje milosti, iskazivanje ljubavi, borbe za istinom, pravdom i slobodom, izricanja zahvala i oprosta, jačanje volja u užem smislu itd. ) njezinih pojedinaca bez obzira na njihova vjerska, životna ili svjetonazorska opredjeljenja.Introducation: A fast is one food and one drink restriction for one certain time. It could be the word about «cancelling» the food for the time or an absolutely restriction of no food or no drink for one day or several days. Fasting is very important for every religion but also in general life in regular conditions. A spiritual willing is not one spiritual status per se because it depends at the physical condition for accepting the spiritual dimension. Aim: The aim of this observation is a fast could be considered as one ordered type of a spirital diet or as a certain type of «a spirituial tablet». Methods: Clients\u27 anamnestic data during interview for their life styles and physiological functions such as their kinds of food consummation before any psychiatric, psychodinamic or hagiotherapeutic treatmant were analysed. Results: In analysed poll could be included only who declared oneself for religious person. But results coud not be reliably because one certain discrepanty is between regular and unregular religious practices and religious statements. For discussion is what one medical doctor or one hagioterapist could recommend about various types of suiting fasts to their clients for periodical or for long time which could not be harmful for their health specially for risky groups as the youth in development or for old people. Conclusion: However in spite of the discussing controled fasting could be good for the general health specially to its spiritual component because it could positive impact on better charity, better love, biger willing for verity, justice, freedom, thankfulness and amnesty and also on stronger volition as one special mechanism etc. without one\u27s life style or religious orientation

The assessment of DNA damage and repair following exposure to sevoflurane in vivo

Cilj istraživanja bio je istražiti povezanost oštećenja i popravka DNA u leukocitima periferne krvi, stanicama jetre, bubrega i mozga Swiss albino miševa uzrokovanog uzastopnim izlaganjem sevofluranu. Miševi su bili izlagani sevofluranu u dozi od 2.4 vol% po dva sata dnevno, 3 dana uzastopce. Za procjenu genetičkog oštećenja spomenutih stanica upotrijebljeni su komet test i mikronukleus test. Za provođenje komet testa miševi su podijeljeni u 5 pokusnih skupina (kontrolna skupina, skupina miševa žrtvovanih treći dan pokusa neposredno nakon izlaganja sevofluranu, 2 sata, 6 sati i 24 sata nakon izlaganja sevofluranu). Rezultati su pokazali statistički značajno povećanje srednje vrijednosti TL, TM i TI u svim pregledanim organima u usporedbi sa kontrolom. Veličina oštećenja DNA u točno određenom vremenu nakon uzastopnog izlaganja sevofluranu, bila je drugačija za svaki pregledani organ. Najveće oštećenje DNA uočeno je odmah nakon izlaganja sevofluranu, posebno u leukocitima periferne krvi. Inducirano oštećenje se dogodilo šest sati kasnije u organima nego u stanicama krvi, što je bilo za očekivati zbog toksikokinetike lijekova, jer se sevofluran prvo apsorbira u krvi. Niti jedno od proučavanih tkiva nije pokazivalo znakove popravka sve dok nisu prošla 24 sata od izlaganja sevofluranu. Za detektiranje nepopravljenih oštećenih dijelova genoma in vivo upotrijebljen je mikronukleus test. Broj mikronukleusa u retikulocitima statistički se značajno povećao u svim proučavanim vremenskim razmacima nakon tretmana u odnosu na kontrolnu skupinu.Aim of the study was to investigate the relationship between DNA damage and repair of peripheral blood leucocytes (PBL), liver, kidney and brain cells of Swiss albino mice induced by repeated exposure to sevoflurane. Mice were exposed to sevoflurane in a dose of 2.4vol% for two hours daily, for three consecutive days. Genetic damage of mentioned cells was investigated using comet assay and micronucleus test. To perform comet assay, mice were divided into 5 experimental groups (control group, groups of exposed mice sacrificed third day of experiment immediately after exposure ot sevoflurane, two hours, six hours and twenty-four hours later). The results showed statistically significant increase in mean TL, TM and TI values in all examined organs vs. the control group. The DNA damage at a specific time after the repeated exposure to sevoflurane was different for each examined organ. Significant DNA damage immediately post exposure to sevoflurane was observed in PBL. The induced damage in organs occured 6 hours later than in PBL what was expected according to toxicokinetic of drugs, where blood is the first compartment to absorb the sevoflurane. However none of the assayed tissues revealed signs of repair until 24th hour after the exposure. To distinguish unrepaired genome damage in vivo micronucleus test was applied. Number of micronuclei in reticulocytes showed statistically significant increase compared with control group at all observed times after the treatment

The assessment of DNA damage and repair following exposure to sevoflurane in vivo

Cilj istraživanja bio je istražiti povezanost oštećenja i popravka DNA u leukocitima periferne krvi, stanicama jetre, bubrega i mozga Swiss albino miševa uzrokovanog uzastopnim izlaganjem sevofluranu. Miševi su bili izlagani sevofluranu u dozi od 2.4 vol% po dva sata dnevno, 3 dana uzastopce. Za procjenu genetičkog oštećenja spomenutih stanica upotrijebljeni su komet test i mikronukleus test. Za provođenje komet testa miševi su podijeljeni u 5 pokusnih skupina (kontrolna skupina, skupina miševa žrtvovanih treći dan pokusa neposredno nakon izlaganja sevofluranu, 2 sata, 6 sati i 24 sata nakon izlaganja sevofluranu). Rezultati su pokazali statistički značajno povećanje srednje vrijednosti TL, TM i TI u svim pregledanim organima u usporedbi sa kontrolom. Veličina oštećenja DNA u točno određenom vremenu nakon uzastopnog izlaganja sevofluranu, bila je drugačija za svaki pregledani organ. Najveće oštećenje DNA uočeno je odmah nakon izlaganja sevofluranu, posebno u leukocitima periferne krvi. Inducirano oštećenje se dogodilo šest sati kasnije u organima nego u stanicama krvi, što je bilo za očekivati zbog toksikokinetike lijekova, jer se sevofluran prvo apsorbira u krvi. Niti jedno od proučavanih tkiva nije pokazivalo znakove popravka sve dok nisu prošla 24 sata od izlaganja sevofluranu. Za detektiranje nepopravljenih oštećenih dijelova genoma in vivo upotrijebljen je mikronukleus test. Broj mikronukleusa u retikulocitima statistički se značajno povećao u svim proučavanim vremenskim razmacima nakon tretmana u odnosu na kontrolnu skupinu.Aim of the study was to investigate the relationship between DNA damage and repair of peripheral blood leucocytes (PBL), liver, kidney and brain cells of Swiss albino mice induced by repeated exposure to sevoflurane. Mice were exposed to sevoflurane in a dose of 2.4vol% for two hours daily, for three consecutive days. Genetic damage of mentioned cells was investigated using comet assay and micronucleus test. To perform comet assay, mice were divided into 5 experimental groups (control group, groups of exposed mice sacrificed third day of experiment immediately after exposure ot sevoflurane, two hours, six hours and twenty-four hours later). The results showed statistically significant increase in mean TL, TM and TI values in all examined organs vs. the control group. The DNA damage at a specific time after the repeated exposure to sevoflurane was different for each examined organ. Significant DNA damage immediately post exposure to sevoflurane was observed in PBL. The induced damage in organs occured 6 hours later than in PBL what was expected according to toxicokinetic of drugs, where blood is the first compartment to absorb the sevoflurane. However none of the assayed tissues revealed signs of repair until 24th hour after the exposure. To distinguish unrepaired genome damage in vivo micronucleus test was applied. Number of micronuclei in reticulocytes showed statistically significant increase compared with control group at all observed times after the treatment